US20170081705A1 - Microfluidic Analysis System - Google Patents
Microfluidic Analysis System Download PDFInfo
- Publication number
- US20170081705A1 US20170081705A1 US15/278,894 US201615278894A US2017081705A1 US 20170081705 A1 US20170081705 A1 US 20170081705A1 US 201615278894 A US201615278894 A US 201615278894A US 2017081705 A1 US2017081705 A1 US 2017081705A1
- Authority
- US
- United States
- Prior art keywords
- sample
- carrier fluid
- nucleic acid
- acid molecules
- thermal
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000004458 analytical method Methods 0.000 title claims abstract description 16
- 239000012530 fluid Substances 0.000 claims abstract description 69
- 238000003752 polymerase chain reaction Methods 0.000 claims abstract description 18
- 239000000523 sample Substances 0.000 claims description 61
- 238000000034 method Methods 0.000 claims description 20
- 238000001514 detection method Methods 0.000 claims description 4
- 230000003287 optical effect Effects 0.000 claims description 4
- 239000000376 reactant Substances 0.000 claims description 4
- 210000001124 body fluid Anatomy 0.000 claims description 2
- 150000007523 nucleic acids Chemical class 0.000 claims 13
- 102000039446 nucleic acids Human genes 0.000 claims 13
- 108020004707 nucleic acids Proteins 0.000 claims 13
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 abstract description 2
- 239000000470 constituent Substances 0.000 abstract description 2
- 229910052710 silicon Inorganic materials 0.000 abstract description 2
- 239000010703 silicon Substances 0.000 abstract description 2
- 239000003921 oil Substances 0.000 abstract 1
- 229920002545 silicone oil Polymers 0.000 description 7
- 238000005382 thermal cycling Methods 0.000 description 6
- 206010028980 Neoplasm Diseases 0.000 description 5
- 230000003321 amplification Effects 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 238000011109 contamination Methods 0.000 description 4
- 230000002068 genetic effect Effects 0.000 description 4
- 238000003199 nucleic acid amplification method Methods 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 201000011510 cancer Diseases 0.000 description 3
- 230000035772 mutation Effects 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 230000008901 benefit Effects 0.000 description 2
- 239000012620 biological material Substances 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 238000004088 simulation Methods 0.000 description 2
- 239000012472 biological sample Substances 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 230000002759 chromosomal effect Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000001351 cycling effect Effects 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 210000004602 germ cell Anatomy 0.000 description 1
- 230000005484 gravity Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005459 micromachining Methods 0.000 description 1
- 230000008450 motivation Effects 0.000 description 1
- 230000000771 oncological effect Effects 0.000 description 1
- 238000005086 pumping Methods 0.000 description 1
- 238000007670 refining Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 230000000392 somatic effect Effects 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F25/00—Flow mixers; Mixers for falling materials, e.g. solid particles
- B01F25/10—Mixing by creating a vortex flow, e.g. by tangential introduction of flow components
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F33/00—Other mixers; Mixing plants; Combinations of mixers
- B01F33/30—Micromixers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/50273—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the means or forces applied to move the fluids
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502769—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by multiphase flow arrangements
- B01L3/502776—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by multiphase flow arrangements specially adapted for focusing or laminating flows
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502769—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by multiphase flow arrangements
- B01L3/502784—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by multiphase flow arrangements specially adapted for droplet or plug flow, e.g. digital microfluidics
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L7/00—Heating or cooling apparatus; Heat insulating devices
- B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
- B01L7/525—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples with physical movement of samples between temperature zones
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
- G01N35/08—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/06—Fluid handling related problems
- B01L2200/0636—Focussing flows, e.g. to laminate flows
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/06—Fluid handling related problems
- B01L2200/0673—Handling of plugs of fluid surrounded by immiscible fluid
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/14—Process control and prevention of errors
- B01L2200/141—Preventing contamination, tampering
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0809—Geometry, shape and general structure rectangular shaped
- B01L2300/0816—Cards, e.g. flat sample carriers usually with flow in two horizontal directions
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
- B01L2300/087—Multiple sequential chambers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/18—Means for temperature control
- B01L2300/1838—Means for temperature control using fluid heat transfer medium
- B01L2300/185—Means for temperature control using fluid heat transfer medium using a liquid as fluid
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/04—Moving fluids with specific forces or mechanical means
- B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
- B01L2400/0409—Moving fluids with specific forces or mechanical means specific forces centrifugal forces
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/04—Moving fluids with specific forces or mechanical means
- B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
- B01L2400/0487—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
- G01N2035/00465—Separating and mixing arrangements
- G01N2035/00514—Stationary mixing elements
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
- G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
- G01N35/1095—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices for supplying the samples to flow-through analysers
Definitions
- the invention relates to analysis systems for analysis such as Polymerase Chain Reaction (PCR) analysis to detect the population of rare mutated cells in a sample of bodily fluid and/or tissue.
- PCR Polymerase Chain Reaction
- a common method is to probe the sample using known genetic markers, the markers being specific to the type of mutation being sought, and then amplify the targets in the sample. If the mutations or chromosomal aberations are present then the amplification can be detected, usually using optical techniques.
- PCR Polymerase Chain Reaction
- 6,306,590 describes a method of performing a PCR in a microfluidic device, in which a channel heats, and then cools PCR reactants cyclically.
- U.S. Pat. No. 6,670,153 also describes use of a microfluidic device for PCR.
- the invention is directed towards providing an improved microfluidic analysis system for applications such as the above.
- a biological sample analysis system comprising:
- the analysis stages comprise a thermal cycling stage and an optical detection stage for performance of a polymerise chain reaction.
- the sample preparation stage comprises a centrifuge for separation of samples from an input fluid and for introduction of the samples to the primary carrier fluid.
- the centrifuge comprises a pair of opposed primary carrier fluid channels on either side of a vortex chamber, whereby flow of primary carrier fluid in said channels causes centrifuging of sample in the vortex chamber and flow of sample from the chamber into said channels.
- contact between the sample and the vortex chamber surface is avoided by wrapping the sample in an initial carrier fluid within the chamber.
- the controller directs separation in the centrifuge either radially or axially due to gravity according to nature of the input fluid such as blood containing the sample.
- the primary carrier fluid velocity is in the range of 1 m/s to 20 m/s.
- the thermal cycling stage comprises a microfluidic thermal device comprising a thermal zone comprising a sample inlet for flow of sample through a sample channel while enveloped in the primary carrier fluid, and a thermal carrier inlet for flow of a thermal carrier fluid to heat or cool the sample by heat conduction through the primary carrier fluid.
- microfluidic thermal device thermal zone further comprises separate sample and thermal outlets positioned to allow flow of thermal carrier fluid into and out of contact with the primary carrier fluid.
- the thermal cycling stage comprises a plurality of thermal zones.
- the microfluidic thermal device comprises a plurality of thermal zones in series.
- the thermal cycling stage comprises a plurality of microfluidic thermal devices in series.
- the microfluidic thermal device comprises a closed sample channel for re-circulation of sample with successive heating or cooling in successive thermal zones.
- the controller directs flow of the thermal and primary carrier fluids to control flowrate of sample by enveloping within the primary carrier fluid and by viscous drag between the thermal carrier fluid and the primary carrier fluid.
- the primary carrier fluid is biologically non-reactive.
- the primary carrier fluid is a silicone oil.
- the thermal carrier fluid is biologically non-reactive.
- the thermal carrier fluid is a silicone oil.
- the temperatures and flowrates of the carrier fluids are controlled to achieve a temperature ramping gradient of 17° C./sec to 25° C./sec.
- FIG. 1 is a diagram of an analysis system of the invention.
- FIG. 2 is a diagrammatic plan view of a centrifuge of the system, and FIG. 3 is a simulation diagram showing centrifuging;
- FIG. 4 is a perspective view of the main body of a microfluidic beater of the system
- FIG. 5 is a prediction velocity and temperature plot along a thermal stage of the heater
- FIG. 6 is a centre line temperature profile in the flow direction showing fast response of same in the heated zone.
- FIG. 7 is a plan view of an alternative microfluidic heater.
- an analysis system 1 comprises a controller 2 which interfaces with various stages.
- a carrier fluid supply 4 delivers carrier fluid to a macro pump 5 which delivers it at a high flowrate to a sample preparation stage 6 .
- the latter also receives a bio-fluid sample, and centrifuges the sample in a vortex created by carrier fluid flow, as described in more detail below.
- Reactants are supplied by a supply 8 to a flow controller 7 which delivers streams of separated DNA with reactants enveloped in carrier fluid to a thermal cycling stage 9 .
- the DNA is amplified in the stage 9 and optically detected by a detection stage 10 .
- the samples are enveloped in a biologically non-reactive carrier fluid such as silicone oil. This avoids risk of contamination from residual molecules on system channel surfaces.
- a centrifuge device 20 of the sample preparation stage 6 is illustrated diagrammatically. It comprises opposed carrier supply lines 21 and 22 and a central vortex chamber 23 having a sample inlet out of the plane of the page.
- the centrifuge 20 operates by primary carrier fluid in the channels 21 and 22 driving sample fluid in the chamber 23 into a vortex via viscous forces at the interface between the two fluids.
- the carrier fluid is silicone oil mixed to be neutrally buoyant with the sample.
- FIG. 3 illustrates the centrifuging activity, the greater density of dots indicating higher flow velocities.
- the left-hand scale shows the velocity range of 1 m/s to 20 m/s.
- the sample is wrapped in an initial volume of carrier fluid within the chamber 23 to prevent surface contamination.
- the carrier fluid is pumped at speeds of 5 ms ⁇ 1 through the system.
- the desired carrier fluid speed is 1 m/s to 20 m/s.
- the device has further potential to be miniaturized to centrifuge at up to 200,000 g, as these levels of force are necessary for efficient separation of RNA and other smaller cellular constituents and bio-molecules.
- the continuous throughput centrifuge offers many benefits over conventional technology.
- the device may also function as a fluid mixing device by reversing the flow path of one of the carrier fluid, if such is desired for an application. It is modular in nature, meaning two or more systems can be placed together in any configuration and run by the same control and power source system.
- the centrifuge 20 has no moving parts thereby allowing excellent reliability compared with a system having moving pans. An important consequence of this feature is that manufacturing this device at the micro-scale using current silicon processing or micro-machining is readily achievable.
- a microfluidic thermal device 51 of the stage 9 is shown. It comprises three successive thermal zones 52 , 53 , and 54 . Each zone comprises a sample inlet 60 and an outlet 61 for flow of the bio sample in the primary carrier fluid. There are also a pair of thermal carrier inlets 65 and 66 , and a pair of thermal carrier outlets 67 and 68 for each of the three zones.
- This drawing shows only the main body, there also being top and bottom sealing transparent plates.
- the bio sample which enters the sample inlet 60 of each stage is enveloped and conveyed by the carrier fluid henceforth called the “primary carrier fluid”.
- Thermal carrier fluid is delivered at the inlets 65 and 66 to heat or cool the bio sample via the primary carrier fluid.
- the arrangement of a number (in this case three) of thermal zones in series offers advantages to applications such as the polymerase chain reaction (PCR) where rapid and numerous thermal cycles lead to dramatic amplification of a DNA template strand.
- PCR polymerase chain reaction
- the device 51 also acts as an ejector pump, in which the velocity and hence the residency time of the sample is controlled by controlling velocity of one or both of the carriers fluids.
- the carrier flow parameters determine how long the sample remains at the set temperature in each zone. This is often important, as chemical reactions require particular times for completion.
- the device 51 can therefore be tuned to the required residency times and ramp rates by controlling the carrier velocity.
- a predicted velocity contour map at the mid-height plane of a zone channel is shown.
- Carrier fluid enters through the channels at the top and bottom left of the image and exits through the channels at the top and bottom right of the image.
- the sample fluid enters and exits through the central channel.
- the different shadings of this map indicate the velocities, the range being 0.01 m/s to 0.1 m/s.
- sample fluid enters through the central channel at the left of the image at a temperature of 50° C. and is heated to 70° C. by the thermal carrier fluid.
- FIG. 6 shows a temperature profile along a longitudinal centerline of a thermal zone.
- a target temperature of 342 K is achieved within an extremely short distance from entrance, achieving an excellent temperature ramp rate of 20° C./sec over a distance of 0.05 m.
- a ramping of 17° C./sec to 25° C./sec is desirable for many applications.
- FIG. 7 another microfluidic thermal device, 70 , is shown.
- the zones 71 and 73 are on one side and there is only a single zone, 72 , on the other side.
- the thermal carrier fluid is silicone oil, as is the primary carrier fluid.
- the thermal carrier fluid for the zone 71 is at 68° C., to ramp up the bio sample to this temperature during residency in this zone.
- the zones 72 and 73 provide outlet temperatures of 95° C. and 72° C. respectively.
- the optical detection stage 10 is positioned over the microfluidic device 70 to analyse the sample.
- the silicone oil is sufficiently transparent to detect the fluorescently tagged molecules.
- the invention achieves comprehensive control over bio sample flowrate and temperature, with no risk of contamination from device surfaces.
- the invention also achieves integrated pumping and thermal cycling of the sample without moving parts at the microscale. There are very high throughputs as measured by processing time for one sample.
- the system is expected to have a low cost and high reliability due to the absence of micro scale moving parts.
- the system also allows independent control and variation of all PCR parameters for process optimisation.
Abstract
Description
- This is a CONTINUATION of PCT/IE2004/000115 filed 6 Sep. 2004 and published in English, claiming the priorities of U.S. Application No. 60/500,344 and 60/500,345, both filed on 5 Sep. 2003.
- The invention relates to analysis systems for analysis such as Polymerase Chain Reaction (PCR) analysis to detect the population of rare mutated cells in a sample of bodily fluid and/or tissue.
- Prior Art Discussion
- It is known for at least the past decade that cancers have a genetic cause. With the emergence of fast methods of sequencing and the publication of the human genome, the motivation and methods are available to find the genetic causes, both germline and somatic, of the most prevalent cancers. Contemporary oncological research suggests that there is a sequence of mutations that must occur for a cancer to be life-threatening, called the multistage model. Cancer could therefore be diagnosed earlier by detecting these genetic markers thereby increasing the probability of cure. However, even with refining of the sample, the target cells and their DNA are still usually very rare, perhaps one part in 106. The analysis system must therefore be able to perform very effective amplification.
- There are several methods of attempting to identify rare cells in a sample of bio-fluid. A common method is to probe the sample using known genetic markers, the markers being specific to the type of mutation being sought, and then amplify the targets in the sample. If the mutations or chromosomal aberations are present then the amplification can be detected, usually using optical techniques.
- It is also possible, depending on the amplification used, to use the Polymerase Chain Reaction (PCR) to detect the number of mutated cells in the original sample: a number important as firstly, it can be linked to the progress of the cancer and secondly, it provides a quantitative measure with which to diagnose remission. PCR is the enzyme-catalysed reaction used to amplify the sample. It entails taking a small quantity of DNA or RNA and producing many identical copies of it in vitro. A system to achieve a. PCR is to process the samples by thermally cycling them is described in U.S. Pat. No. 5,270,183. However, this apparently involves a risk of sample contamination by surfaces in the temperature zones and other channels. Also, U.S. Pat. No. 6,306,590 describes a method of performing a PCR in a microfluidic device, in which a channel heats, and then cools PCR reactants cyclically. U.S. Pat. No. 6,670,153 also describes use of a microfluidic device for PCR.
- The invention is directed towards providing an improved microfluidic analysis system for applications such as the above.
- According to the invention, there is provided a biological sample analysis system comprising:
-
- a carrier fluid;
- a sample supply;
- a sample preparation stage for providing a flow of sample enveloped in a primary carrier fluid;
- at least one analysis stage for performing analysis of the sample while controlling flow of the sample while enveloped within the primary carrier fluid without the sample contacting a solid surface; and a controller for controlling the system.
- In one embodiment, the analysis stages comprise a thermal cycling stage and an optical detection stage for performance of a polymerise chain reaction.
- In another embodiment, the sample preparation stage comprises a centrifuge for separation of samples from an input fluid and for introduction of the samples to the primary carrier fluid.
- In a further embodiment, the centrifuge comprises a pair of opposed primary carrier fluid channels on either side of a vortex chamber, whereby flow of primary carrier fluid in said channels causes centrifuging of sample in the vortex chamber and flow of sample from the chamber into said channels.
- In one embodiment, contact between the sample and the vortex chamber surface is avoided by wrapping the sample in an initial carrier fluid within the chamber.
- In another embodiment, the controller directs separation in the centrifuge either radially or axially due to gravity according to nature of the input fluid such as blood containing the sample.
- In a further embodiment, the primary carrier fluid velocity is in the range of 1 m/s to 20 m/s.
- In one embodiment, the thermal cycling stage comprises a microfluidic thermal device comprising a thermal zone comprising a sample inlet for flow of sample through a sample channel while enveloped in the primary carrier fluid, and a thermal carrier inlet for flow of a thermal carrier fluid to heat or cool the sample by heat conduction through the primary carrier fluid.
- In another embodiment, the microfluidic thermal device thermal zone further comprises separate sample and thermal outlets positioned to allow flow of thermal carrier fluid into and out of contact with the primary carrier fluid.
- In a further embodiment, there is at least one pair of opposed thermal carrier inlet/outlet pairs on opposed sides of a sample channel.
- In one embodiment, the thermal cycling stage comprises a plurality of thermal zones.
- In one embodiment, the microfluidic thermal device comprises a plurality of thermal zones in series.
- In another embodiment, the thermal cycling stage comprises a plurality of microfluidic thermal devices in series.
- In a further embodiment, the microfluidic thermal device comprises a closed sample channel for re-circulation of sample with successive heating or cooling in successive thermal zones.
- In one embodiment, the controller directs flow of the thermal and primary carrier fluids to control flowrate of sample by enveloping within the primary carrier fluid and by viscous drag between the thermal carrier fluid and the primary carrier fluid.
- In another embodiment, the primary carrier fluid is biologically non-reactive.
- In a further embodiment, the primary carrier fluid is a silicone oil.
- In one embodiment, the thermal carrier fluid is biologically non-reactive.
- In another embodiment, the thermal carrier fluid is a silicone oil.
- In a further embodiment, the temperatures and flowrates of the carrier fluids are controlled to achieve a temperature ramping gradient of 17° C./sec to 25° C./sec.
- The invention will be more clearly understood from the following description of some embodiments thereof, given by way of example only with reference to the accompanying drawings in which:
-
FIG. 1 is a diagram of an analysis system of the invention. -
FIG. 2 is a diagrammatic plan view of a centrifuge of the system, andFIG. 3 is a simulation diagram showing centrifuging; -
FIG. 4 is a perspective view of the main body of a microfluidic beater of the system; -
FIG. 5 is a prediction velocity and temperature plot along a thermal stage of the heater; -
FIG. 6 is a centre line temperature profile in the flow direction showing fast response of same in the heated zone; and -
FIG. 7 is a plan view of an alternative microfluidic heater. - Referring to
FIG. 1 an analysis system 1 comprises a controller 2 which interfaces with various stages. A carrier fluid supply 4 delivers carrier fluid to a macro pump 5 which delivers it at a high flowrate to a sample preparation stage 6. The latter also receives a bio-fluid sample, and centrifuges the sample in a vortex created by carrier fluid flow, as described in more detail below. Reactants are supplied by a supply 8 to a flow controller 7 which delivers streams of separated DNA with reactants enveloped in carrier fluid to a thermal cycling stage 9. The DNA is amplified in the stage 9 and optically detected by adetection stage 10. Throughout the process the samples are enveloped in a biologically non-reactive carrier fluid such as silicone oil. This avoids risk of contamination from residual molecules on system channel surfaces. - Referring to
FIG. 2 a centrifuge device 20 of the sample preparation stage 6 is illustrated diagrammatically. It comprises opposedcarrier supply lines central vortex chamber 23 having a sample inlet out of the plane of the page. The centrifuge 20 operates by primary carrier fluid in thechannels chamber 23 into a vortex via viscous forces at the interface between the two fluids. In this embodiment, the carrier fluid is silicone oil mixed to be neutrally buoyant with the sample. - The vortex, or centrifuge, is thus established without any mechanical moving parts. The carrier fluid drives a vortex of the sample to be centrifuged thereby avoiding the very many difficulties of designing and operating moving parts at the micro scale, particularly at high rotational speeds.
FIG. 3 illustrates the centrifuging activity, the greater density of dots indicating higher flow velocities. The left-hand scale shows the velocity range of 1 m/s to 20 m/s. The sample is wrapped in an initial volume of carrier fluid within thechamber 23 to prevent surface contamination. - This achieves a continuous throughput micro-centrifuging to suitably extract DNA and RNA from cellular material. The bio-fluid is centrifuged resulting in DNA and other bio-molecules of interest accumulating at the bottom of the chamber, thereby providing an efficient and simple method of manipulating micron and sub-micron quantities of bio-fluid. The DNA and RNA are separated due to the greater weight and viscous resistance of the DNA. Numerical simulations (
FIG. 3 ) of the flow show that tangential velocities of up to 10 ms−1 are generated towards the edge of the vortex core. Calculations reveal this to be equivalent to a rotational speed of almost 20,000 rpm or 2,000 g in terms of a centrifugal force. In order to achieve these levels of centrifugal force, the carrier fluid is pumped at speeds of 5 ms−1 through the system. In general, the desired carrier fluid speed is 1 m/s to 20 m/s. The device has further potential to be miniaturized to centrifuge at up to 200,000 g, as these levels of force are necessary for efficient separation of RNA and other smaller cellular constituents and bio-molecules. - Overall, the continuous throughput centrifuge offers many benefits over conventional technology. The device may also function as a fluid mixing device by reversing the flow path of one of the carrier fluid, if such is desired for an application. It is modular in nature, meaning two or more systems can be placed together in any configuration and run by the same control and power source system. The centrifuge 20 has no moving parts thereby allowing excellent reliability compared with a system having moving pans. An important consequence of this feature is that manufacturing this device at the micro-scale using current silicon processing or micro-machining is readily achievable.
- Referring to
FIG. 4 a microfluidicthermal device 51 of the stage 9 is shown. It comprises three successivethermal zones sample inlet 60 and anoutlet 61 for flow of the bio sample in the primary carrier fluid. There are also a pair ofthermal carrier inlets thermal carrier outlets - The bio sample which enters the
sample inlet 60 of each stage is enveloped and conveyed by the carrier fluid henceforth called the “primary carrier fluid”. Thermal carrier fluid is delivered at theinlets - As the sample remains in a low shear rate region of the flow, mass transport by diffusion of sample species is kept to a minimum. The low shear region reduces damage by shear to macro molecules that may be carried by the bio sample. The arrangement of a number (in this case three) of thermal zones in series offers advantages to applications such as the polymerase chain reaction (PCR) where rapid and numerous thermal cycles lead to dramatic amplification of a DNA template strand.
- The
device 51 also acts as an ejector pump, in which the velocity and hence the residency time of the sample is controlled by controlling velocity of one or both of the carriers fluids. The carrier flow parameters determine how long the sample remains at the set temperature in each zone. This is often important, as chemical reactions require particular times for completion. Thedevice 51 can therefore be tuned to the required residency times and ramp rates by controlling the carrier velocity. - Referring to
FIG. 5 a predicted velocity contour map at the mid-height plane of a zone channel is shown. Carrier fluid enters through the channels at the top and bottom left of the image and exits through the channels at the top and bottom right of the image. The sample fluid enters and exits through the central channel. The different shadings of this map indicate the velocities, the range being 0.01 m/s to 0.1 m/s. - In one example, sample fluid enters through the central channel at the left of the image at a temperature of 50° C. and is heated to 70° C. by the thermal carrier fluid.
-
FIG. 6 shows a temperature profile along a longitudinal centerline of a thermal zone. A target temperature of 342 K is achieved within an extremely short distance from entrance, achieving an excellent temperature ramp rate of 20° C./sec over a distance of 0.05 m. In general, a ramping of 17° C./sec to 25° C./sec is desirable for many applications. - The following table sets out parameters for one example. A silicone oil, density matched to the density of the bio sample, is used for both of the carrier fluids.
-
TABLE 1 Boundary Conditions and Fluid Properties Overall Channel Dimensions 5 mm × 5 mm × 200 mm Wall Boundary Condition outside of carrier Adiabatic flow interaction zones Heat Transfer Carrier Fluid Inlet temperature 70° C., 90° C., 110° C. for each zone Sample/Transport Carrier Inlet Pressure 0 Pa Heat Transfer Carrier Fluid Inlet Pressure 0.2 Pa Sample/Transport Carrier Outlet Pressure 1.9 Pa Heat Transfer Carrier Fluid Outlet Pressure 1.7 Pa Mass Diffusivity 1.3E−12 m2/s Approximate Temperature Gradient in Zones 20° C./sec - Referring to
FIG. 7 , another microfluidic thermal device, 70, is shown. There are again three thermal zones, however in this case on a generally rectangular closed circuit, withzones zones zone 71 is at 68° C., to ramp up the bio sample to this temperature during residency in this zone. Thezones - The
optical detection stage 10 is positioned over themicrofluidic device 70 to analyse the sample. The silicone oil is sufficiently transparent to detect the fluorescently tagged molecules. - It will be appreciated that the invention achieves comprehensive control over bio sample flowrate and temperature, with no risk of contamination from device surfaces. The invention also achieves integrated pumping and thermal cycling of the sample without moving parts at the microscale. There are very high throughputs as measured by processing time for one sample.
- The system is expected to have a low cost and high reliability due to the absence of micro scale moving parts. The system also allows independent control and variation of all PCR parameters for process optimisation.
- The invention is not limited to the embodiments described but may be varied in construction and detail.
Claims (10)
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US15/278,894 US10676786B2 (en) | 2003-09-05 | 2016-09-28 | Microfluidic analysis system |
US16/892,488 US11807902B2 (en) | 2003-09-05 | 2020-06-04 | Microfluidic analysis system |
Applications Claiming Priority (6)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US50034403P | 2003-09-05 | 2003-09-05 | |
US50034503P | 2003-09-05 | 2003-09-05 | |
PCT/IE2004/000115 WO2005023427A1 (en) | 2003-09-05 | 2004-09-06 | A microfluidic analysis system |
US11/366,524 US7622076B2 (en) | 2003-09-05 | 2006-03-03 | Microfluidic analysis system |
US12/617,286 US20100092987A1 (en) | 2003-09-05 | 2009-11-12 | Microfluidic analysis system |
US15/278,894 US10676786B2 (en) | 2003-09-05 | 2016-09-28 | Microfluidic analysis system |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US12/617,286 Division US20100092987A1 (en) | 2003-09-05 | 2009-11-12 | Microfluidic analysis system |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US16/892,488 Continuation US11807902B2 (en) | 2003-09-05 | 2020-06-04 | Microfluidic analysis system |
Publications (2)
Publication Number | Publication Date |
---|---|
US20170081705A1 true US20170081705A1 (en) | 2017-03-23 |
US10676786B2 US10676786B2 (en) | 2020-06-09 |
Family
ID=34278702
Family Applications (4)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US11/366,524 Active 2026-06-01 US7622076B2 (en) | 2003-09-05 | 2006-03-03 | Microfluidic analysis system |
US12/617,286 Abandoned US20100092987A1 (en) | 2003-09-05 | 2009-11-12 | Microfluidic analysis system |
US15/278,894 Active 2025-09-21 US10676786B2 (en) | 2003-09-05 | 2016-09-28 | Microfluidic analysis system |
US16/892,488 Active 2025-04-19 US11807902B2 (en) | 2003-09-05 | 2020-06-04 | Microfluidic analysis system |
Family Applications Before (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US11/366,524 Active 2026-06-01 US7622076B2 (en) | 2003-09-05 | 2006-03-03 | Microfluidic analysis system |
US12/617,286 Abandoned US20100092987A1 (en) | 2003-09-05 | 2009-11-12 | Microfluidic analysis system |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US16/892,488 Active 2025-04-19 US11807902B2 (en) | 2003-09-05 | 2020-06-04 | Microfluidic analysis system |
Country Status (3)
Country | Link |
---|---|
US (4) | US7622076B2 (en) |
EP (1) | EP1663497B2 (en) |
WO (1) | WO2005023427A1 (en) |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US10967338B2 (en) | 2003-09-05 | 2021-04-06 | Stokes Bio Ltd. | Methods of releasing and analyzing cellular components |
WO2021201819A1 (en) * | 2020-03-30 | 2021-10-07 | Hewlett-Packard Development Company, L.P. | Intermittent warming of a biologic sample including a nucleic acid |
US11772096B2 (en) | 2006-02-07 | 2023-10-03 | Stokes Bio Ltd. | System for processing biological sample |
US11788120B2 (en) | 2017-11-27 | 2023-10-17 | The Trustees Of Columbia University In The City Of New York | RNA printing and sequencing devices, methods, and systems |
Families Citing this family (72)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1663497B2 (en) | 2003-09-05 | 2020-03-25 | Stokes Bio Limited | A microfluidic analysis system |
US8968659B2 (en) | 2003-09-05 | 2015-03-03 | Stokes Bio Limited | Sample dispensing |
US7968287B2 (en) | 2004-10-08 | 2011-06-28 | Medical Research Council Harvard University | In vitro evolution in microfluidic systems |
EP3913375A1 (en) | 2006-01-11 | 2021-11-24 | Bio-Rad Laboratories, Inc. | Microfluidic devices and methods of use in the formation and control of nanoreactors |
US8735169B2 (en) | 2006-02-07 | 2014-05-27 | Stokes Bio Limited | Methods for analyzing agricultural and environmental samples |
US8501497B2 (en) | 2006-02-07 | 2013-08-06 | Stokes Bio Limited | Forming sample combinations using liquid bridge systems |
US20080280331A1 (en) * | 2006-02-07 | 2008-11-13 | Stokes Bio Limited | Microfluidic Analysis System |
US20100304446A1 (en) * | 2006-02-07 | 2010-12-02 | Stokes Bio Limited | Devices, systems, and methods for amplifying nucleic acids |
US9562837B2 (en) | 2006-05-11 | 2017-02-07 | Raindance Technologies, Inc. | Systems for handling microfludic droplets |
US20080014589A1 (en) | 2006-05-11 | 2008-01-17 | Link Darren R | Microfluidic devices and methods of use thereof |
EP2069070B1 (en) * | 2006-09-28 | 2013-11-27 | Stokes Bio Limited | A qpcr analysis apparatus |
US8772046B2 (en) | 2007-02-06 | 2014-07-08 | Brandeis University | Manipulation of fluids and reactions in microfluidic systems |
WO2008116941A1 (en) | 2007-03-26 | 2008-10-02 | Fundación Gaiker | Method and device for detecting genetic material by means of polymerase chain reaction |
WO2008130623A1 (en) | 2007-04-19 | 2008-10-30 | Brandeis University | Manipulation of fluids, fluid components and reactions in microfluidic systems |
EP4047367A1 (en) | 2008-07-18 | 2022-08-24 | Bio-Rad Laboratories, Inc. | Method for detecting target analytes with droplet libraries |
WO2010133965A2 (en) | 2009-05-19 | 2010-11-25 | Life Technologies Corporation | Sampling device |
US9399797B2 (en) | 2010-02-12 | 2016-07-26 | Raindance Technologies, Inc. | Digital analyte analysis |
EP3392349A1 (en) | 2010-02-12 | 2018-10-24 | Raindance Technologies, Inc. | Digital analyte analysis |
US9366632B2 (en) | 2010-02-12 | 2016-06-14 | Raindance Technologies, Inc. | Digital analyte analysis |
US10351905B2 (en) | 2010-02-12 | 2019-07-16 | Bio-Rad Laboratories, Inc. | Digital analyte analysis |
EP3447155A1 (en) | 2010-09-30 | 2019-02-27 | Raindance Technologies, Inc. | Sandwich assays in droplets |
EP2673382B1 (en) * | 2011-02-11 | 2020-05-06 | Bio-Rad Laboratories, Inc. | Thermocycling device for nucleic acid amplification and methods of use |
WO2012109600A2 (en) | 2011-02-11 | 2012-08-16 | Raindance Technologies, Inc. | Methods for forming mixed droplets |
WO2012112804A1 (en) | 2011-02-18 | 2012-08-23 | Raindance Technoligies, Inc. | Compositions and methods for molecular labeling |
CN103917293B (en) | 2011-04-08 | 2016-01-20 | 斯多克斯生物有限公司 | Biological detection system and using method |
WO2012139041A1 (en) | 2011-04-08 | 2012-10-11 | Stokes Bio Limited | System and method for charging fluids |
EP3709018A1 (en) | 2011-06-02 | 2020-09-16 | Bio-Rad Laboratories, Inc. | Microfluidic apparatus for identifying components of a chemical reaction |
US8658430B2 (en) | 2011-07-20 | 2014-02-25 | Raindance Technologies, Inc. | Manipulating droplet size |
US9855559B2 (en) | 2011-12-30 | 2018-01-02 | Abbott Molecular Inc. | Microorganism nucleic acid purification from host samples |
US10323279B2 (en) | 2012-08-14 | 2019-06-18 | 10X Genomics, Inc. | Methods and systems for processing polynucleotides |
US11591637B2 (en) | 2012-08-14 | 2023-02-28 | 10X Genomics, Inc. | Compositions and methods for sample processing |
MX364957B (en) | 2012-08-14 | 2019-05-15 | 10X Genomics Inc | Microcapsule compositions and methods. |
US10400280B2 (en) | 2012-08-14 | 2019-09-03 | 10X Genomics, Inc. | Methods and systems for processing polynucleotides |
US9701998B2 (en) | 2012-12-14 | 2017-07-11 | 10X Genomics, Inc. | Methods and systems for processing polynucleotides |
US10533221B2 (en) | 2012-12-14 | 2020-01-14 | 10X Genomics, Inc. | Methods and systems for processing polynucleotides |
WO2014124338A1 (en) | 2013-02-08 | 2014-08-14 | 10X Technologies, Inc. | Polynucleotide barcode generation |
US11901041B2 (en) | 2013-10-04 | 2024-02-13 | Bio-Rad Laboratories, Inc. | Digital analysis of nucleic acid modification |
US9944977B2 (en) | 2013-12-12 | 2018-04-17 | Raindance Technologies, Inc. | Distinguishing rare variations in a nucleic acid sequence from a sample |
US9824068B2 (en) | 2013-12-16 | 2017-11-21 | 10X Genomics, Inc. | Methods and apparatus for sorting data |
CN106795553B (en) | 2014-06-26 | 2021-06-04 | 10X基因组学有限公司 | Methods of analyzing nucleic acids from individual cells or cell populations |
US10221436B2 (en) | 2015-01-12 | 2019-03-05 | 10X Genomics, Inc. | Processes and systems for preparation of nucleic acid sequencing libraries and libraries prepared using same |
US10647981B1 (en) | 2015-09-08 | 2020-05-12 | Bio-Rad Laboratories, Inc. | Nucleic acid library generation methods and compositions |
US11371094B2 (en) | 2015-11-19 | 2022-06-28 | 10X Genomics, Inc. | Systems and methods for nucleic acid processing using degenerate nucleotides |
US11081208B2 (en) | 2016-02-11 | 2021-08-03 | 10X Genomics, Inc. | Systems, methods, and media for de novo assembly of whole genome sequence data |
US10815525B2 (en) | 2016-12-22 | 2020-10-27 | 10X Genomics, Inc. | Methods and systems for processing polynucleotides |
US10550429B2 (en) | 2016-12-22 | 2020-02-04 | 10X Genomics, Inc. | Methods and systems for processing polynucleotides |
EP4029939B1 (en) | 2017-01-30 | 2023-06-28 | 10X Genomics, Inc. | Methods and systems for droplet-based single cell barcoding |
US10995333B2 (en) | 2017-02-06 | 2021-05-04 | 10X Genomics, Inc. | Systems and methods for nucleic acid preparation |
US10544413B2 (en) | 2017-05-18 | 2020-01-28 | 10X Genomics, Inc. | Methods and systems for sorting droplets and beads |
EP3625353B1 (en) | 2017-05-18 | 2022-11-30 | 10X Genomics, Inc. | Methods and systems for sorting droplets and beads |
US20190064173A1 (en) | 2017-08-22 | 2019-02-28 | 10X Genomics, Inc. | Methods of producing droplets including a particle and an analyte |
US10837047B2 (en) | 2017-10-04 | 2020-11-17 | 10X Genomics, Inc. | Compositions, methods, and systems for bead formation using improved polymers |
WO2019083852A1 (en) | 2017-10-26 | 2019-05-02 | 10X Genomics, Inc. | Microfluidic channel networks for partitioning |
WO2019084043A1 (en) | 2017-10-26 | 2019-05-02 | 10X Genomics, Inc. | Methods and systems for nuclecic acid preparation and chromatin analysis |
CN111479631B (en) | 2017-10-27 | 2022-02-22 | 10X基因组学有限公司 | Methods and systems for sample preparation and analysis |
EP3954782A1 (en) | 2017-11-15 | 2022-02-16 | 10X Genomics, Inc. | Functionalized gel beads |
WO2019108851A1 (en) | 2017-11-30 | 2019-06-06 | 10X Genomics, Inc. | Systems and methods for nucleic acid preparation and analysis |
WO2019157529A1 (en) | 2018-02-12 | 2019-08-15 | 10X Genomics, Inc. | Methods characterizing multiple analytes from individual cells or cell populations |
US11639928B2 (en) | 2018-02-22 | 2023-05-02 | 10X Genomics, Inc. | Methods and systems for characterizing analytes from individual cells or cell populations |
US11471884B2 (en) | 2018-04-02 | 2022-10-18 | Dropworks, Inc. | Systems and methods for serial flow emulsion processes |
WO2019195166A1 (en) | 2018-04-06 | 2019-10-10 | 10X Genomics, Inc. | Systems and methods for quality control in single cell processing |
US11703427B2 (en) | 2018-06-25 | 2023-07-18 | 10X Genomics, Inc. | Methods and systems for cell and bead processing |
US20200032335A1 (en) | 2018-07-27 | 2020-01-30 | 10X Genomics, Inc. | Systems and methods for metabolome analysis |
US11459607B1 (en) | 2018-12-10 | 2022-10-04 | 10X Genomics, Inc. | Systems and methods for processing-nucleic acid molecules from a single cell using sequential co-partitioning and composite barcodes |
US11845983B1 (en) | 2019-01-09 | 2023-12-19 | 10X Genomics, Inc. | Methods and systems for multiplexing of droplet based assays |
US11851683B1 (en) | 2019-02-12 | 2023-12-26 | 10X Genomics, Inc. | Methods and systems for selective analysis of cellular samples |
WO2020168013A1 (en) | 2019-02-12 | 2020-08-20 | 10X Genomics, Inc. | Methods for processing nucleic acid molecules |
US11467153B2 (en) | 2019-02-12 | 2022-10-11 | 10X Genomics, Inc. | Methods for processing nucleic acid molecules |
US11655499B1 (en) | 2019-02-25 | 2023-05-23 | 10X Genomics, Inc. | Detection of sequence elements in nucleic acid molecules |
US11920183B2 (en) | 2019-03-11 | 2024-03-05 | 10X Genomics, Inc. | Systems and methods for processing optically tagged beads |
US11851700B1 (en) | 2020-05-13 | 2023-12-26 | 10X Genomics, Inc. | Methods, kits, and compositions for processing extracellular molecules |
CN113607586A (en) * | 2021-07-19 | 2021-11-05 | 国网浙江省电力有限公司经济技术研究院 | Drop-wall flow separation test device |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20040180346A1 (en) * | 2003-03-14 | 2004-09-16 | The Regents Of The University Of California. | Chemical amplification based on fluid partitioning |
Family Cites Families (71)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
FR2572180B1 (en) | 1984-10-24 | 1987-03-20 | Eric Marteau D Autry | METHOD AND APPARATUS FOR REPAIRING SAMPLES FOR ANALYSIS |
US5102517A (en) | 1990-05-23 | 1992-04-07 | Millipore Corporation | Capillary wash system |
US5641622A (en) | 1990-09-13 | 1997-06-24 | Baxter International Inc. | Continuous centrifugation process for the separation of biological components from heterogeneous cell populations |
US5270183A (en) | 1991-02-08 | 1993-12-14 | Beckman Research Institute Of The City Of Hope | Device and method for the automated cycling of solutions between two or more temperatures |
ATE208658T1 (en) | 1993-07-28 | 2001-11-15 | Pe Corp Ny | APPARATUS AND METHOD FOR NUCLEIC ACID DUPLICATION |
US6143496A (en) | 1997-04-17 | 2000-11-07 | Cytonix Corporation | Method of sampling, amplifying and quantifying segment of nucleic acid, polymerase chain reaction assembly having nanoliter-sized sample chambers, and method of filling assembly |
DE19736691A1 (en) * | 1997-08-22 | 1999-02-25 | Michael Prof Dr Med Giesing | Characterising and identifying disseminated metastatic cancer cells |
WO1999041015A1 (en) * | 1998-02-11 | 1999-08-19 | Institut für Physikalische Hochtechnologie e.V. | Miniaturized temperature-zone flow reactor |
US6306590B1 (en) | 1998-06-08 | 2001-10-23 | Caliper Technologies Corp. | Microfluidic matrix localization apparatus and methods |
US6261431B1 (en) * | 1998-12-28 | 2001-07-17 | Affymetrix, Inc. | Process for microfabrication of an integrated PCR-CE device and products produced by the same |
US6294063B1 (en) * | 1999-02-12 | 2001-09-25 | Board Of Regents, The University Of Texas System | Method and apparatus for programmable fluidic processing |
US20020182749A1 (en) | 1999-05-11 | 2002-12-05 | Aclara Biosciences, Inc. | Sample evaporative control |
US6193471B1 (en) | 1999-06-30 | 2001-02-27 | Perseptive Biosystems, Inc. | Pneumatic control of formation and transport of small volume liquid samples |
US6524456B1 (en) | 1999-08-12 | 2003-02-25 | Ut-Battelle, Llc | Microfluidic devices for the controlled manipulation of small volumes |
US6355164B1 (en) * | 1999-10-29 | 2002-03-12 | Ontogen Corporation | Sample collection apparatus and method for multiple channel high throughput purification |
US6481453B1 (en) | 2000-04-14 | 2002-11-19 | Nanostream, Inc. | Microfluidic branch metering systems and methods |
WO2001089691A2 (en) | 2000-05-24 | 2001-11-29 | Micronics, Inc. | Capillaries for fluid movement within microfluidic channels |
WO2001089696A2 (en) | 2000-05-24 | 2001-11-29 | Micronics, Inc. | Microfluidic concentration gradient loop |
US20010048637A1 (en) | 2000-05-24 | 2001-12-06 | Weigl Bernhard H. | Microfluidic system and method |
DE10055318A1 (en) | 2000-06-09 | 2001-12-20 | Advalytix Ag | Process for specific directed manipulation of small amounts of materials on solid body surfaces comprises producing an impulse along the solid body surface, and causing the impulse |
ATE448875T1 (en) | 2000-09-14 | 2009-12-15 | Caliper Life Sciences Inc | MICROFLUIDIC DEVICES AND METHODS FOR CARRYING OUT TEMPERATURE-MEDIATED REACTIONS |
EP1334347A1 (en) | 2000-09-15 | 2003-08-13 | California Institute Of Technology | Microfabricated crossflow devices and methods |
GB0026404D0 (en) | 2000-10-28 | 2000-12-13 | Siddall & Hilton Ltd | Body support arrangements |
EP1343973B2 (en) | 2000-11-16 | 2020-09-16 | California Institute Of Technology | Apparatus and methods for conducting assays and high throughput screening |
WO2002072264A1 (en) | 2001-03-09 | 2002-09-19 | Biomicro Systems, Inc. | Method and system for microfluidic interfacing to arrays |
CA2442978A1 (en) | 2001-04-04 | 2002-10-17 | Bioprocessors Corp. | System and method for dispensing liquids |
US7077152B2 (en) | 2001-07-07 | 2006-07-18 | Nanostream, Inc. | Microfluidic metering systems and methods |
AUPR707101A0 (en) * | 2001-08-16 | 2001-09-06 | Corbett Research Pty Ltd | Continuous flow thermal device |
US6907895B2 (en) | 2001-09-19 | 2005-06-21 | The United States Of America As Represented By The Secretary Of Commerce | Method for microfluidic flow manipulation |
US20030073089A1 (en) | 2001-10-16 | 2003-04-17 | Mauze Ganapati R. | Companion cartridge for disposable diagnostic sensing platforms |
US7189580B2 (en) | 2001-10-19 | 2007-03-13 | Wisconsin Alumni Research Foundation | Method of pumping fluid through a microfluidic device |
US20030138819A1 (en) | 2001-10-26 | 2003-07-24 | Haiqing Gong | Method for detecting disease |
WO2003057010A2 (en) | 2002-01-04 | 2003-07-17 | Board Of Regents, The University Of Texas System | Droplet-based microfluidic oligonucleotide synthesis engine |
US7718099B2 (en) | 2002-04-25 | 2010-05-18 | Tosoh Corporation | Fine channel device, fine particle producing method and solvent extraction method |
FR2839504B1 (en) * | 2002-05-07 | 2004-06-18 | Commissariat Energie Atomique | DEVICE AND METHOD FOR DISPENSING LIQUID PRODUCTS |
EP2282214B1 (en) | 2002-05-09 | 2022-10-05 | The University of Chicago | Device and method for pressure-driven plug transport and reaction |
US7901939B2 (en) | 2002-05-09 | 2011-03-08 | University Of Chicago | Method for performing crystallization and reactions in pressure-driven fluid plugs |
GB2395196B (en) | 2002-11-14 | 2006-12-27 | Univ Cardiff | Microfluidic device and methods for construction and application |
US7547380B2 (en) * | 2003-01-13 | 2009-06-16 | North Carolina State University | Droplet transportation devices and methods having a fluid surface |
GB0315438D0 (en) | 2003-07-02 | 2003-08-06 | Univ Manchester | Analysis of mixed cell populations |
US7767435B2 (en) * | 2003-08-25 | 2010-08-03 | University Of Washington | Method and device for biochemical detection and analysis of subcellular compartments from a single cell |
EP1663497B2 (en) | 2003-09-05 | 2020-03-25 | Stokes Bio Limited | A microfluidic analysis system |
US9597644B2 (en) | 2003-09-05 | 2017-03-21 | Stokes Bio Limited | Methods for culturing and analyzing cells |
US8968659B2 (en) | 2003-09-05 | 2015-03-03 | Stokes Bio Limited | Sample dispensing |
JP4341372B2 (en) | 2003-10-30 | 2009-10-07 | コニカミノルタホールディングス株式会社 | Liquid mixing method, mixing apparatus and mixing system |
US7687269B2 (en) * | 2003-12-10 | 2010-03-30 | Northeastern University | Method for efficient transport of small liquid volumes to, from or within microfluidic devices |
WO2005080606A1 (en) | 2004-02-18 | 2005-09-01 | Xiaochuan Zhou | Fluidic devices and methods for multiplex chemical and biochemical reactions |
KR100552706B1 (en) | 2004-03-12 | 2006-02-20 | 삼성전자주식회사 | Method and apparatus for nucleic acid amplification |
US20050221339A1 (en) | 2004-03-31 | 2005-10-06 | Medical Research Council Harvard University | Compartmentalised screening by microfluidic control |
US20050272144A1 (en) | 2004-06-08 | 2005-12-08 | Konica Minolta Medical & Graphic, Inc. | Micro-reactor for improving efficiency of liquid mixing and reaction |
US7655470B2 (en) | 2004-10-29 | 2010-02-02 | University Of Chicago | Method for manipulating a plurality of plugs and performing reactions therein in microfluidic systems |
WO2006089192A2 (en) | 2005-02-18 | 2006-08-24 | Canon U.S. Life Sciences, Inc. | Devices and methods for identifying genomic dna of organisms |
US20070014695A1 (en) | 2005-04-26 | 2007-01-18 | Applera Corporation | Systems and Methods for Multiple Analyte Detection |
EP2660482B1 (en) | 2005-08-22 | 2019-08-07 | Life Technologies Corporation | Vorrichtung, System und Verfahren unter Verwendung von nichtmischbaren Flüssigkeiten mit unterschiedlichen Volumen |
US20070134209A1 (en) * | 2005-12-12 | 2007-06-14 | Metafluidics, Inc. | Cellular encapsulation for self-assembly of engineered tissue |
US20080280331A1 (en) | 2006-02-07 | 2008-11-13 | Stokes Bio Limited | Microfluidic Analysis System |
US8735169B2 (en) | 2006-02-07 | 2014-05-27 | Stokes Bio Limited | Methods for analyzing agricultural and environmental samples |
US8501497B2 (en) | 2006-02-07 | 2013-08-06 | Stokes Bio Limited | Forming sample combinations using liquid bridge systems |
EP1981625B1 (en) | 2006-02-07 | 2010-08-18 | Stokes Bio Limited | A microfluidic droplet queuing network and method |
ATE523244T1 (en) | 2006-02-07 | 2011-09-15 | Stokes Bio Ltd | LIQUID BRIDGE SYSTEM AND METHOD |
US8492168B2 (en) | 2006-04-18 | 2013-07-23 | Advanced Liquid Logic Inc. | Droplet-based affinity assays |
US20080014589A1 (en) | 2006-05-11 | 2008-01-17 | Link Darren R | Microfluidic devices and methods of use thereof |
EP2069070B1 (en) | 2006-09-28 | 2013-11-27 | Stokes Bio Limited | A qpcr analysis apparatus |
US8202686B2 (en) | 2007-03-22 | 2012-06-19 | Advanced Liquid Logic, Inc. | Enzyme assays for a droplet actuator |
WO2008130623A1 (en) | 2007-04-19 | 2008-10-30 | Brandeis University | Manipulation of fluids, fluid components and reactions in microfluidic systems |
EP4047367A1 (en) * | 2008-07-18 | 2022-08-24 | Bio-Rad Laboratories, Inc. | Method for detecting target analytes with droplet libraries |
US20100059120A1 (en) | 2008-09-11 | 2010-03-11 | General Electric Company | Microfluidic device and methods for droplet generation and manipulation |
US8697011B2 (en) | 2009-05-19 | 2014-04-15 | Stokes Bio Limited | Sampling device with immiscible fluid supply tube in counter-flow arrangement |
US9625454B2 (en) | 2009-09-04 | 2017-04-18 | The Research Foundation For The State University Of New York | Rapid and continuous analyte processing in droplet microfluidic devices |
EP3392349A1 (en) | 2010-02-12 | 2018-10-24 | Raindance Technologies, Inc. | Digital analyte analysis |
CA2946144A1 (en) | 2014-04-21 | 2015-10-29 | President And Fellows Of Harvard College | Systems and methods for barcoding nucleic acids |
-
2004
- 2004-09-06 EP EP04770390.5A patent/EP1663497B2/en active Active
- 2004-09-06 WO PCT/IE2004/000115 patent/WO2005023427A1/en active Application Filing
-
2006
- 2006-03-03 US US11/366,524 patent/US7622076B2/en active Active
-
2009
- 2009-11-12 US US12/617,286 patent/US20100092987A1/en not_active Abandoned
-
2016
- 2016-09-28 US US15/278,894 patent/US10676786B2/en active Active
-
2020
- 2020-06-04 US US16/892,488 patent/US11807902B2/en active Active
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20040180346A1 (en) * | 2003-03-14 | 2004-09-16 | The Regents Of The University Of California. | Chemical amplification based on fluid partitioning |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US10967338B2 (en) | 2003-09-05 | 2021-04-06 | Stokes Bio Ltd. | Methods of releasing and analyzing cellular components |
US11772096B2 (en) | 2006-02-07 | 2023-10-03 | Stokes Bio Ltd. | System for processing biological sample |
US11788120B2 (en) | 2017-11-27 | 2023-10-17 | The Trustees Of Columbia University In The City Of New York | RNA printing and sequencing devices, methods, and systems |
WO2021201819A1 (en) * | 2020-03-30 | 2021-10-07 | Hewlett-Packard Development Company, L.P. | Intermittent warming of a biologic sample including a nucleic acid |
Also Published As
Publication number | Publication date |
---|---|
EP1663497B1 (en) | 2014-08-27 |
US20200354772A1 (en) | 2020-11-12 |
US20100092987A1 (en) | 2010-04-15 |
US20060205062A1 (en) | 2006-09-14 |
WO2005023427A1 (en) | 2005-03-17 |
US7622076B2 (en) | 2009-11-24 |
EP1663497B2 (en) | 2020-03-25 |
EP1663497A1 (en) | 2006-06-07 |
US11807902B2 (en) | 2023-11-07 |
US10676786B2 (en) | 2020-06-09 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US11807902B2 (en) | Microfluidic analysis system | |
US10543466B2 (en) | High resolution temperature profile creation in a digital microfluidic device | |
Yuen et al. | Microchip module for blood sample preparation and nucleic acid amplification reactions | |
US10239057B2 (en) | Microfluidic devices and methods for cell analysis and molecular diagnostics | |
AU2008222590B2 (en) | Apparatus and method for nucleic acid amplification | |
EP0637999B1 (en) | Polynucleotide amplification analysis using a microfabricated device | |
KR102127231B1 (en) | Plurality Of Reaction Chambers In A Test Cartridge | |
WO2004073863A2 (en) | Chemical reactions apparatus | |
US20120045765A1 (en) | Composite liquid cells | |
US20070190641A1 (en) | Mesoscale polynucleotide amplification device and method | |
Lien et al. | Miniaturization of molecular biological techniques for gene assay | |
WO2005082535A1 (en) | Buoyancy-driven microfluidics | |
KR100579831B1 (en) | Microfluidic sample processing apparatus capable of assembling | |
Kye et al. | Separation, purification, and detection of cfDNA in a microfluidic device | |
KR102043103B1 (en) | Automatic genetic detector | |
IE20040589A1 (en) | A microfluidic analysis system | |
IE84099B1 (en) | A microfluidic analysis system | |
US20230008992A1 (en) | Devices for generating pre-templated instant partitions | |
Brahmasandra et al. | A microfabricated fluidic reaction and separation system for integrated DNA analysis | |
US20160375441A1 (en) | Apparatus and method for nucleic acid amplification | |
CN116083223A (en) | Nucleic acid detection equipment and system | |
IE20050100A1 (en) | Microfluidics | |
IE84261B1 (en) | Microfluidics | |
WO2014113663A1 (en) | In-line polymerase chain reaction |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AS | Assignment |
Owner name: STOKES BIO LTD., IRELAND Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:DAVIES, MARK;DALTON, TARA;REEL/FRAME:046583/0095 Effective date: 20091208 |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: NON FINAL ACTION MAILED |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: RESPONSE TO NON-FINAL OFFICE ACTION ENTERED AND FORWARDED TO EXAMINER |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: NON FINAL ACTION MAILED |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: RESPONSE TO NON-FINAL OFFICE ACTION ENTERED AND FORWARDED TO EXAMINER |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: NOTICE OF ALLOWANCE MAILED -- APPLICATION RECEIVED IN OFFICE OF PUBLICATIONS |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: PUBLICATIONS -- ISSUE FEE PAYMENT VERIFIED |
|
STCF | Information on status: patent grant |
Free format text: PATENTED CASE |
|
MAFP | Maintenance fee payment |
Free format text: PAYMENT OF MAINTENANCE FEE, 4TH YEAR, LARGE ENTITY (ORIGINAL EVENT CODE: M1551); ENTITY STATUS OF PATENT OWNER: LARGE ENTITY Year of fee payment: 4 |